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Benzyl-N-acetyl-α-d-galactosaminide inhibits the sialylation and the secretion of mucins by a mucin secreting HT-29 cell subpopulation

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Abstract

We have analysed the mucins synthesized by the HT-29 MTX cell subpopulation, derived from the HT-29 human colon carcinoma cells through a selective pressure with methotrexate (Lesuffleuret al., 1990,Cancer Res 50: 6334–43), in the presence of benzyl-N-acetyl-α-galactosaminide (GalNAcα-O-benzyl), which is a potential competitive inhibitor of the β1,3-galactosyltransferase that synthesizes the T-antigen. The main observation was a 13-fold decrease in the sialic acid content of mucins after 24 h of exposure to 5mm GalNAcα-O-benzyl. This effect was accompanied by an increased reactivity of these mucins to peanut lectin, testifying to the higher amount of T-antigen. The second observation was a decrease in the secretion of the mucins by GalNAcα-O-benzyl treated cells. The decrease in mucin sialyation was achieved through thein situ β-galactosylation of GalNAcα-O-benzyl into Galβ1–3GalNAcα-O-benzyl, which acts as a competitive substrate of Galβ1–3GalNAc α2,3-sialyltransferase, as shown by the intracellular accumulation of NeuAcα2–3Galβ1–3GalNAcα-O-benzyl in treated cells.

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Abbreviations

BSM:

bovine submaxillary mucin

MTX:

methotrexate

PBS:

sodium phosphate 10mm, NaCl 0.15m, pH 7.4 buffer

pNp:

p-nitrophenol

TBS:

Tris/HCl 10mm, NaCl 0.15m, pH 7.4 buffer

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Authors and Affiliations

  1. Laboratoire de Chimie Biologique, Unité Mixte de Recherche du CNRS no 111, Université des Sciences et Technologies de Lille, F-59655, Villeneuve d'Ascq, France

    Philippe Delannoy & Andre Verbert

  2. INSERM U-377, place de Verdun, F-59045, Lille, France

    Isabelle Kim, Nathalie Emery, Pierre Degand & Guillemette Huet

  3. Institut Municipal d'Investigacio Medica, Universitat Autonoma de Barcelona, Spain

    Carmen de Bolos

Authors
  1. Philippe Delannoy
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  2. Isabelle Kim
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  3. Nathalie Emery
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  4. Carmen de Bolos
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  5. Andre Verbert
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  6. Pierre Degand
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  7. Guillemette Huet
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Additional information

Enzymes: CMP-NeuAc: Galβ1–3/4GlcNAc α2,3-sialyltransferase, ST3(N), EC 2.4.99.6; CMP-NeuAc: Galβ1–4GlcNAc α2,6-sialyltransferase, ST6(N), EC 2.4.99.1; CMP-NeuAc: Galβ1–3GalNAc α2,3-sialyltransferase, ST3(O), EC 2.4.99.4; CMP-NeuAc: R-GalNAcα1-O-Ser α2,6-sialyltransferase, ST6(O)-I, EC 2.4.99.3; CMP-NeuAc: NeuAcα2–3Galβ1–3GalNAc α2,6-sialyltransferase, ST6(O)-II, EC 2.4.99.7; UDP-GlcNAc: Galβ1–3GalNAc-R·(GlcNAc to GalNAc) β1,6-N-acetylglucosaminyltransferase, EC 2.4.1.102; UDP-GlcNAc: GalNAcα-R β1,3-N-acetylglucosaminyltransferase, EC 2.4.1.147; UDP-Gal: GalNAc-R β1,3-galactosyltransferase, EC 2.4.1.122.

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Delannoy, P., Kim, I., Emery, N. et al. Benzyl-N-acetyl-α-d-galactosaminide inhibits the sialylation and the secretion of mucins by a mucin secreting HT-29 cell subpopulation. Glycoconjugate J 13, 717–726 (1996). https://doi.org/10.1007/BF00702335

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  • DOI: https://doi.org/10.1007/BF00702335

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